Anticancer Effect of Deuterium Oxide on a Bladder Cancer Cell Related to Bcl-2 and Bax

Jong Yoon Bahk; Jeong-Hee Lee; Hong Suk Chung; Hae Young Lee; Bong Chul Chung; Moon Seok Park; Seung Ki Min; Myeong Ok Kim

Journal of Industrial and Engineering Chemistry, Vol.13, No.4, 501-507, July, 2007

Jong Yoon Bahk, Jeong-Hee Lee¹, Hong Suk Chung², Hae Young Lee³, Bong Chul Chung^{4, †}, Moon Seok Park³, Seung Ki Min⁵, and Myeong Ok Kim^{3, †}

Department of Urology, Korea
¹Pathology, Medical School, Gyeongsang National University, Jinju, 660-751, Korea
²Korea Atomic Energy Research Institute, PO Box 105, Daejeon, 305-600, Korea
³Division of Life Science and Applied of Life Science, BK 21 College of Natural Sciences, Gyeongsang NationalUniversity, Jinju, 660-701, Korea
⁴Bioanalysis and Biotransformation Research Center, Korea Institute of Science and Technology, P.O.BOX 131,Seoul, Korea
⁵Department of Urology, National Police Hospital, Seoul, 138-708, Korea

E-mail:,

To evaluate the potentiality, as a drug for an intravesical instillation after a transurethral resection of a bladder tumor, we studied the anticancer effects of deuterium oxide (D2O) related to bcl-2 and bax. Bladder cancer cell T-24 was used and culture media were prepared with H2O and D2O at different concentrations (D2O v/v), 0 (control), 75, and 100 %. Cells were exposed to each D2O for 2, 2.5, 3, and 3.5 h. The antiproliferative effects were measured by a quantitative colorimetric assay (MTT assay) and a hemocytometer. Invasion study was implemented with a modified reconstituted basement membrane after an exposure to D2O. Immunohistochemical staining and Western blot analysis for bcl-2 and bax were implemented to evaluate the relation between D2O and apoptosis. Marked cytoreduction was observed at an exposure to both 75 and 100 % of D2O. The cellular proliferation in 75 % D2O was 68.95, 46.77, 24.65, and 14.41 % when compared to the control at an exposure time of 2, 2.5, 3, and 3.5 h, respectively. The comparative ratios in 100 % were 37.93, 4.30, 1.00, and 0.18 % at each exposure group. An invasion of the D2O exposed cells into the reconstituted basement membrane did not occur. Morphologic cellular findings suggested an apoptosis. The bcl-2 and bax proteins were noted from most cells regardless of an exposure to D2O. The expressions of the bcl-2 were decreased with an increased exposure time in both the 75 and 100 % D2O. The expressions of the bax were increased with an increased exposure time for both concentrations. D2O has high antiproliferative, and anti-invasive effects at 75 and 100 % D2O with an exposure time longer than 2.5 h. We concluded that the mechanism of the antiproliferative effect of the D2O would be mediated by an apoptosis. And the cytotoxic results related to D2O show the potential for an application as an agent for a post TUR-BT intravesical instillation in a superficial bladder cancer.

Keywords:antiproliferation;deuterium oxide;bladder cancer cell

[References]

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Melekos MD, Chionis H, Pantazakos A, Fokaefs E, Paranychianakis G, Dauaher H, J. Urol., 149, 744 (1993)
Pashos CL, Botteman MF, Laskin BL, Redaelli A, Cancer Pract., 10, 311 (2002)
Yoshimura K, Utsunomiya N, Ichioka K, Matsui Y, Terai A, Arai Y, Urology, 65, 290 (2005)
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Altermatt HJ, Gebbers JO, Laissue JA, Cancer, 62, 462 (1988)
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Hodel A, Gebbers JO, Cottier H, Laissue JA, Life Sci., 30, 1987 (1982)
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Thrasher JB, Crawford ED, Urol. Clin. North Am., 19, 529 (1992)

[Related Articles]

[1] Heney NM, Ahmed S, Flanagan MJ, Frable W, Corder MP, Hafermann MD, Hawkins IR, J. Urol., 130, 1083 (1983)

[2] Torti FM, Lum BL, J. Clin. Oncol., 2, 505 (1984)

[3] Soloway MS, Masters S, Cancer, 46, 1158 (1980)

[4] Pode D, Alon Y, Horowitz AT, Vlodavsky I, Biran S, J. Urol., 136, 482 (1986)

[5] Igawa M, Urakami S, Shirakawa H, Shiina H, Ishibe T, Usui T, Moriyama H, Hiroshima J. Med. Sci., 44, 93 (1995)

[6] Solomon LZ, Jennings AM, Foley SJ, Birch BR, Cooper AJ, Br. J. Urol., 82, 122 (1998)

[7] Cheng L, Gu J, Ulbright TM, MacLennan GT, Sweeney CJ, Zhang S, Sanchez K, Koch MO, Eble JN, Cancer, 94, 104 (2002)

[8] Melekos MD, Chionis H, Pantazakos A, Fokaefs E, Paranychianakis G, Dauaher H, J. Urol., 149, 744 (1993)

[9] Pashos CL, Botteman MF, Laskin BL, Redaelli A, Cancer Pract., 10, 311 (2002)

[10] Yoshimura K, Utsunomiya N, Ichioka K, Matsui Y, Terai A, Arai Y, Urology, 65, 290 (2005)

[11] McLaughlin JK, Silverman DT, Hsing AW, Ross RK, Schoenberg JB, Yu MC, Stemhagen A, Lynch CF, Blot WJ, Fraumeni JF, Cancer Res., 52, 254 (1992)

[12] Sorahan T, Lancashire RJ, Sole G, Br. J. Urol., 74, 753 (1994)

[13] Altermatt HJ, Gebbers JO, Laissue JA, Cancer, 62, 462 (1988)

[14] Takeda H, Nio Y, Omori H, Uegaki K, Hirahara N, Sasaki S, Tamura K, Ohtani H, Anti-Cancer Drugs, 9, 715 (1998)

[15] Uemura T, Moritake K, Akiyama Y, Kimura Y, Shingu T, Yamasaki T, J Neurosurg., 96, 900 (2002)

[16] Hodel A, Gebbers JO, Cottier H, Laissue JA, Life Sci., 30, 1987 (1982)

[17] Gross PR, Spindel W, Science, 131, 37 (1960)

[18] Lamprecht J, Schroeter D, Paweletz N, Eur. J. Cell Biol., 51, 303 (1990)

[19] Peppard RJ, Karn CA, McCabe MA, Wassall SR, Liechty EA, Denne SC, J. Pediatr., 123, 457 (1993)

[20] Norberg A, Sandhagen B, Bratteby LE, Gabrielsson J, Jones AW, Fan H, Hahn RG, Alcohol Clin. Exp. Res., 25, 1423 (2001)

[21] Oltvai ZN, Milliman CL, Llorsmeyer SJ, Cell, 74, 609 (1993)

[22] Gazzaniga P, Gradilone A, Vercillo R, Gandini O, Silvestri I, Napolitano M, Albonici L, Vincenzoni A, Gallucci M, Frati L, Agliano AM, Int. J. Cancer, 69, 100 (1996)

[23] Korkolopoulou P, Lazaris AP, Konstantinidou AE, Kavantzas N, Patsouris E, Christodoulou P, Thomas-Tsagli E, Davaris P, Eur. Urol., 41, 274 (2002)

[24] Joo HN, Lee CG, J. Ind. Eng. Chem., 13(1), 110 (2007)

[25] Lee WK, Im SM, J. Ind. Eng. Chem., 12(6), 932 (2006)

[26] Reed JC, Hematol. Oncol. Clin. North Am., 9, 451 (1995)

[27] Miyake H, Hanada N, Nakamura H, Kagawa S, Fujiwara T, Hara I, Eto H, Gohji K, Arakawa S, Kamidono S, Saya H, Oncogene, 16, 933 (1998)

[28] Boersma AW, Nooter K, Burger H, Kortland CJ, Stoter G, Cytometry, 27, 275 (1997)

[29] Omori H, Nio Y, Taloda AH, Tamur K, Gan To Kagoku Ryoho, 23, 1665 (1996)

[30] Kushner DJ, Baker A, Dunstall TG, Can. J. Physiol. Pharmacol., 77, 79 (1999)

[31] Coward WA, Cole TJ, Sawyer MB, Prentice AM, Hum. Nutr. Clin. Nutr., 36, 141 (1982)

[32] Park MS, Hahn CH, Chung H, La LY, Kim MO, Bahk JY, J. Ind. Eng. Chem., 9(6), 640 (2003)

[33] Yang B, Gu F, Wang X, Zhonghua Wai Ke Za Zhi, 35, 602 (1997)

[34] Gazzaniga P, Gradilone A, Silvestri I, Gandini O, Giuliani L, Vincenzoni A, Gallucci M, Frati L, Agliano AM, Oncol. Rep., 5, 901 (1998)

[35] Malkowicz SB, Management of superficial bladder cancer: In: Campbell’s urology, 8th ed. Edited by P.C. Walsh, A. B. Retik, E. D. Vaughan, Jr. and A. J. Wein, and W. B. Saunders Co., chapt. 77, pp 2785-2801, Philadelphia (2002)

[36] Thrasher JB, Crawford ED, Urol. Clin. North Am., 19, 529 (1992)